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Blastomere fate and embryonic genome activation (EGA) during human embryonic development are unsolved areas of high scientific and clinical interest. Forty-nine blastomeres from 5- to 8-cell human embryos have been investigated following an efficient single-cell cDNA amplification protocol to provide a template for high-density microarray analysis. The previously described markers, characteristic of Inner Cell Mass (ICM) (n = 120), stemness (n = 190) and Trophectoderm (TE) (n = 45), were analyzed, and a housekeeping pattern of 46 genes was established. All the human blastomeres from the 5- to 8-cell stage embryo displayed a common gene expression pattern corresponding to ICM markers (e.g.,  DDX3 ,  FOXD3 ,  LEFTY1 ,  MYC ,  NANOG ,  POU5F1 ), stemness (e.g.,  POU5F1 ,  DNMT3B ,  GABRB3 ,  SOX2 ,  ZFP42 ,  TERT ), and TE markers (e.g.,  GATA6 ,  EOMES ,  CDX2 ,  LHCGR ). The EGA profile was also investigated between the 5-6- and 8-cell stage embryos, and compared to the blastocyst stage. Known genes (n = 92) such as depleted maternal transcripts (e.g.,  CCNA1 ,  CCNB1 ,  DPPA2 ) and embryo-specific activation (e.g.,  POU5F1 ,  CDH1 ,  DPPA4 ), as well as novel genes, were confirmed. In summary, the global single-cell cDNA amplification microarray analysis of the 5- to 8-cell stage human embryos reveals that blastomere fate is not committed to ICM or TE. Finally, new EGA features in human embryogenesis are presented.

Functional Genomics of 5- to 8-Cell Stage Human Embryos by Blastomere Single-Cell cDNA Analysis